Induction of Various Mutations during PCRs with Manganese and 8-Hydroxy-dGTP
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概要
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To induce various mutations randomly, PCRs with Mn2+ and with a mutagenic deoxyribonucleotide, 8-hydroxy-dGTP (8-OH-dGTP), were performed. Mutations were induced by deoxyribonucleotide imbalance plus 500mM Mn2+ in the Mn2+ PCR, and the amplified DNA was inserted into a plasmid. The plasmid library obtained from the transformed bacterial cells was then used as the template in the next PCR, which was done with 50 or 100mM 8-OH-dGTP. Four kinds of mutations, A:T→G:C and G:C→A:T transitions and A:T→T:A and A:T→C:G transversions, occurred with similar frequencies. These results suggest that this strategy will be useful in random PCR mutagenesis for the in vitro evolution of nucleic acids and proteins, and for analyses of residues in these biomolecules.
- 公益社団法人日本薬学会の論文
- 2007-04-01
著者
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Kamiya Hiroyuki
Faculty of Pharmaceutical Sciences, Hokkaido University
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Harashima Hideyoshi
Faculty of Pharmaceutical Sciences, Hokkaido University
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原島 秀吉
北大院・薬学研究科
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Ito M
Meijo Univ. Nagoya Jpn
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Ito M
Graduate School Of Pharmaceutical Sciences Kyoto University
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Ito M
Department Of Pharmacognosy Graduate School Of Pharmaceutical Sciences Kyoto University
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Ito Michiho
北里大学 薬学部
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Shim Chang
Faculty Of Pharmaceutical Sciences University Of Tokyo : (present Address) College Of Pharmacy Seoul
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Kasai Hiroshi
Inst. Of Industrial Ecological Sciences Univ. Of Occupational And Environmental Health 1-1 Iseigaoka
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Harashima Hideyoshi
Department Of Pharmacokinetics And Pharmaceutics Graduate School Of Pharmaceutical Sciences The Univ
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Kamiya Hiroyuki
産業医科大学産業生態科学研究所
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Murata-kamiya Naoko
Inst. Indust. Ecolog. Sci. Univ. Occup. Environ. Hith.
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Ito Mana
Faculty of Pharmaceutical Sciences, Hokkaido University
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Kamiya Hiroyuki
Faculty Of Pharmaceutical Sciences Hokkaido University
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Harashima Hideyoshi
Faculty Of Pharmaceutical Science The University Of Tokushima
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Kaji Hiroshi
Inst. Indust. Ecolog. Sci. Univ. Occup. Environ. Hith.
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Kamiya Hiroyuki
Faculty Of Pharmaceutical Sciences Hokkaido University:crest Japan Science And Technology
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